论文部分内容阅读
目的:探讨化疗药物对肿瘤增殖活性的影响。方法:选择人宫颈癌细胞系Hela分为两组,分别采用MTT比色法分析测定顺铂处理Hela细胞的浓度;免疫组化SP法分别检测Hela细胞中P27蛋白表达;流式细胞仪分析加药前后细胞周期变化及凋亡情况;用IFFM-D型流动式化学发光仪检测细胞的超弱发光强度。结果:顺铂处理Hela细胞48 h的IC50值为3 mg/L,当DDP浓度在3 mg/L以下时,对Hela细胞无明显毒性作用,超过此浓度时,其毒性呈剂量效应关系(P<0.001);流式细胞仪分析细胞周期可见与Hela细胞相比较,Hela+DDP细胞的G2期细胞数增多,而G1、S期的细胞数明显减少(P<0.01);从细胞凋亡检测显示Hela与Hela+DDP相比,细胞凋亡率在不同时间点明显升高,在24 h、48 h、72 h结果分别为(11.4±5.8、21.8±7.9、32.5±11.6)%。免疫组化结果显示Hela+DDP与Hela细胞相比细胞膜上P27蛋白高表达;在10-4mol/L鲁米诺及0.3%的双氧水(H_2O_2)条件下Hela细胞超弱发光强度高于用Hela+DDP细胞(P<0.001)。结论:超弱发光能够快速、准确、有效地反映肿瘤细胞氧化代谢特点和增殖活动,也可用于筛选敏感的化疗药物。
Objective: To investigate the effect of chemotherapeutic drugs on tumor proliferation activity. Methods: The human cervical cancer cell line Hela was divided into two groups. MTT colorimetric assay was used to determine the concentration of cisplatin-treated Hela cells respectively. The expression of P27 protein in Hela cells was detected by immunohistochemistry SP method. Flow cytometry analysis The changes of cell cycle and apoptosis before and after treatment were measured by IFFM-D flow cytometer. Results: The IC50 value of Hela cells treated with cisplatin for 48 h was 3 mg / L. When DDP concentration was under 3 mg / L, there was no obvious toxic effect on Hela cells. When the concentration was above this concentration, the toxicity was dose-dependent (P <0.001). Compared with Hela cells, the cell number of G2 phase in Hela + DDP cells increased and the number of cells in G1 and S phase decreased significantly (P <0.01) The results showed that compared with Hela + DDP group, the apoptosis rate of Hela significantly increased at different time points, and the results were (11.4 ± 5.8,21.8 ± 7.9,32.5 ± 11.6)% at 24 h, 48 h and 72 h, respectively. The results of immunohistochemistry showed that the expression of P27 protein was higher in Hela + DDP cells than in Hela cells. The ultraweak luminescence intensity of Hela cells was higher than that of Hela + DDP cells in the condition of 10-4mol / L luminol and 0.3% H 2 O 2, DDP cells (P <0.001). CONCLUSION: Ultra-weak luminescence can reflect the characteristics of oxidative metabolism and proliferative activity of tumor cells rapidly, accurately and effectively, and can also be used to screen sensitive chemotherapeutic drugs.